Biotechnology and Recombinant DNA Flashcards
What is Gene Cloning?
creating the exact copy or clone of a particular gene or DNA sequence.
How many plasmids do prokaryotes have?
Multiple
Type II restriction enzymes cut where?
Cut DNA at a defined position close to or inside that recognized sequence.
What bases do restriction enzymes frequently recognize?
4- , 6- , or 8- bases.
Blunt ends can match flush, what of sticky ends?
They have to complement each other to match.
Why are restriction enzymes used?
To protect the bacterial DNA from foreign DNA
What are Vectors?
carrying desired genes into a new cell
Colony Selection is better, why?
You will always get one definite outcome.
Shuttle vectors?
vectors that can be selected in at least 2 different species
Religation vs insertion. Which is much more than likely to occur?
Religation.
How do you detect a product in PCR?
SYBR and TaqMan
rtPCR
Reverse transcription PCR; amplifies RNA.
rtPCR used what primers?
Poly AAA
Transformation is?
DNA from the environment and grow infinitely
Transfection is what?
DNA from the environment but added to mammalian cells.
What are the purpose of Calcium chloride and Lithium Chloride for transformation?
Attaches to the outside of the cell, because the membrane is negatively charged. Causes a pore to appear on the surface.
What is Partial Digestion?
random cuts by restriction enzymes.
Average Gene is how much ?
1000 Bp
Genomic Library Constructions
Partial digestion, creating clones with fragments and transforming to bacteria! You want a lot!
cDNA is made from?
mRNA.
Are all genes there in the cDNA library?
No, not all the genes are turned on at the same time. So only the genes that were turned on will be represented.
Human insulin was used for what technology?
rDNA technology
What approaches are used to treat genetic diseases (3)?
- Gene therapy
- Gene silencing
- Gene editing
Disadvantages of Gene therapy?
DNA could be inserted anywhere.
What two ways can you accomplish gene silencing?
- siRNA and 2. microRNA
RISC is what?
RNA induced silencing complex incorporating siRNA or microRNA
What does siRNA recognize?
long dsRNA, viruses, transposons. This is a defense mechanism.
What do you need to accomplish Gene editing?
Cas9, Guide RNA, Piece of DNA
How was the Human genome project accomplished?
Next-generation sequencing
Herbicide “RoundUp” inhibits what?
Inhibits an enzyme in plants that creates aromatic amino acids.
